Nano-sensor array

ABSTRACT

In one embodiment, a method is provided for the manufacture of a nano-sensor array. A base having a sensing region is provided along with a plurality of nano-sensors. Each of the plurality of nano-sensors is formed by: forming a first nanoneedle along a surface of the base, forming a dielectric on the first nanoneedle, and forming a second nanoneedle on the dielectric layer. The first nanoneedle of each sensor has a first end adjacent to the sensing region of the base. The second nanoneedle is separated from the first nanoneedle by the dielectric and has a first end adjacent the first end of the first nanoneedle. The base is provided with a fluidic channel. The plurality of nano-sensors and the fluidic channel are configured and arranged with the first ends proximate the fluidic channel to facilitate sensing of targeted matter in the fluidic channel.

STATEMENT OF GOVERNMENTAL SUPPORT

This invention was made with U.S. Government support under NationalInstitutes of Health Grant Number P01 HG000205. The U.S. Government hascertain rights in this invention.

FIELD OF THE INVENTION

The present invention relates to chemical and biological particledetection.

BACKGROUND OF THE INVENTION

Biosensors are used for detecting the presence of a target chemical orbiological agent in a wide variety of applications, such as detection ofcontaminants in air (e.g., in air quality sensors) or detecting thepresence of proteins and nucleic acids in blood samples or othersamples. A biosensor measures the presence of various chemicalcomponents in a sample contained within a biosensor cell. For example, abiosensor may be used to measure the amount of glucose present in asample of blood.

Some known methods of detection rely on growth cultures. These methodsare based on the ability of pathogenic species to multiply innutrient-rich medium containing selective agents that inhibit the growthof non-target organisms. These detection methods may be used todifferentiate between target and non-target organisms. Althoughsensitive and accurate, these procedures can take as long as severaldays and thus are not useful for organisms that don't grow easily. Othermethods of detection, such as Polymerase Chain Reaction (PCR) andEnzyme-linked Immunosorbant Assay (ELISA), can directly detectpathogen-specific DNA and proteins, respectively, and can be completedin a matter of hours. PCR is extremely sensitive and has been shown todetect as few as 10 or fewer organisms. In comparison, ELISA is lesssensitive but has the ability to detect proteinaceous toxins. Together,these techniques can provide highly sensitive and specific detection ofpathogens and they are currently the standard techniques used in boththe clinics and research laboratories.

Detection of events such as binding of single cells or molecules isgenerally performed using either optical (usually fluorescent) orelectrical detection. Optical techniques are very sensitive and candetect single molecule events but require the attachment of afluorophore molecule to the target. Optical techniques are moresensitive than are thermal or electrical detection because in opticaldetection, excitement from a single or few photons, in a process such aselectron multiplying or Avalanche phenomena, can be amplified and issufficient to generate a detectable flow of electrons or charge(current). The ability of a microscope to view simultaneously a largearea may be the key. While the fluorescent platforms usually have highersensitivity and signal to noise ratio (SNR) compared to electricalbiosensors, they may not provide real time monitoring possible withelectrical biosensors.

Electrical detection method techniques may not require the attachment offluorophores or other labels but are less sensitive. The devices usuallysuffer from low signal to noise ratio (SNR) due to different sources ofnoise (e.g. electrical, thermal, Flicker, Johnson, etc) and lowdetection signal (e.g. the signal generated due to a reaction or bindingevent of a target molecule to the probe molecule is not large enough).As a result, electrical biosensors exhibit lower sensitivity, incomparison to the some optical detection techniques, which can bedetrimental to early stage detection and diagnosis.

One or more embodiments may address one or more of the above issues.

SUMMARY

Various aspects of one or more embodiments are directed to nano-sensorsand the fabrication thereof.

In one embodiment, a method is provided for the manufacture of anano-sensor array. A base is provided along with a plurality ofnano-sensors. Each of the plurality of nano-sensors is formed by forminga first nanoneedle along a surface of the base, forming a dielectric onthe first nanoneedle, and forming a second nanoneedle on the dielectriclayer. The first nanoneedle of each sensor has a first end adjacent to aregion of the base. The second nanoneedle is separated from the firstnanoneedle by the dielectric and has a first end adjacent the first endof the first nanoneedle. The plurality of nano-sensors and the fluidicchannel are configured and arranged with the first ends in the region tofacilitate sensing of targeted matter in the region.

In another embodiment, a nano-sensor array is provided. The nano-sensorarray includes a base layer and a plurality of nano-sensors. Eachnano-sensor includes a first nanoneedle along a surface of the base, adielectric layer on the first nanoneedle, and a second nanoneedle on thedielectric layer. The first nanoneedle has a first end in a region. Thesecond nanoneedle is separated from the first nanoneedle by thedielectric and has a first end adjacent to the first end of the firstnanoneedle. A probe may be provided on an exposed portion of thedielectric between the first and second nanoneedles at the first end ofthe second nanoneedle. The probe is configured and arranged to bind toparticles of a target type that are present in the fluidic channel. Adetection circuit is coupled to a second end of each of saidnanoneedles. The detection circuit is configured and arranged to detecta change of impedance between the first and second nanoneedles of eachnano-sensor that may result from the binding of a particle of the targettype to the respective probe.

In yet another embodiment, a method of detecting the presence of targetparticles in a sample is provided. The sample is received in a fluidicchannel of a base. One or more target particles in the sample are boundto a probe attached to one or more nano-sensors. Each of the one or morenano-sensors includes a first nanoneedle along a surface of the base, adielectric layer on the first nanoneedle, and a second nanoneedle on thedielectric layer. The first nanoneedle has a first end within thefluidic channel. The second nanoneedle is separated from the firstnanoneedle by the dielectric and has a first end adjacent to the firstend of the first nanoneedle. The probe is located on an exposed portionof the dielectric between the first and second nanoneedles at the firstend of the second nanoneedle. A change of impedance between the firstand second nanoneedles of the one or more nano-sensors is detected. Thechange impedance results from the binding of one of the one or moretarget particles to the probe of the nano-sensor.

It will be appreciated that various other embodiments are set forth inthe Detailed Description and Claims, which follow.

BRIEF DESCRIPTION OF THE DRAWINGS

Various aspects and advantages of the disclosed embodiments will becomeapparent upon review of the following detailed description and uponreference to the drawings in which:

FIG. 1 shows a 3-dimensional view of a nano-sensor array having aplurality of horizontal nano-sensors;

FIG. 2 a shows a top view of a second nano-sensor array having aplurality of horizontal nano-sensors;

FIG. 2 b shows cross section 1 of the nano-sensor array shown in FIG. 2a;

FIG. 2 c shows cross section 2 of the nano-sensor array shown in FIG. 2a;

FIG. 2 d shows cross section 3 of the nano-sensor array shown in FIG. 2a;

FIG. 3 shows a cross section of one implementation of a horizontalnanoneedle-sensor that may be used to implement a nano-sensor array;

FIGS. 4 a shows a circuit diagram of an electrical model of thehorizontal nanoneedle-sensor shown in FIG. 3;

FIG. 4 b shows a circuit diagram of an electrical model of the impedancebetween endpoints A and B of the horizontal nanoneedle-sensor shown inFIG. 3;

FIG. 4 c shows a circuit diagram illustrating the electrical behavior ofthe horizontal nanoneedle-sensor shown in FIG. 3 resulting from in afirst particle binding scenario;

FIG. 4 d shows a circuit diagram illustrating the electrical behavior ofthe horizontal nanoneedle-sensor shown in FIG. 3 resulting from a secondparticle binding scenario;

FIG. 5 a shows a top view of a third nano-sensor array having aplurality of horizontal nano-sensors;

FIG. 5 b shows cross section 1 of the nano-sensor array shown in FIG. 5a;

FIG. 5 c shows cross section 2 of the nano-sensor array shown in FIG. 5a;

FIG. 5 d shows cross section 3 of the nano-sensor array shown in FIG. 5a;

FIG. 6 illustrates a cross-section view of an alternative embodiment ofthe nanoneedle structure shown in FIG. 2 d, wherein ends of nano-sensorsare flush with the wall of a channel formed in the base;

FIG. 7 illustrates a cross-section view of an alternative embodiment ofthe nanoneedle structure shown in FIG. 2 d, wherein the channel isimplemented by sidewalls formed above the base; and

FIG. 8 illustrates the structure and the micrograph of a single and ofdouble nanoneedles.

DETAILED DESCRIPTION

In one or more embodiments, a cost effective nano-sensor array isprovided having a plurality of horizontal nano-sensors having sensingends formed above a channel region of a base. The sensing ends of thenano-sensors are configured to bind with target particles present in asample present in the channel. The unique structure of the horizontalnano-sensors exhibits a change in impedance between two terminals of thenano-sensor in response to the binding of target particles. Using thisstructure, the presence of target particles of interest may beelectrically detected with high sensitivity by measuring the impedanceof the sensors. In this manner, target particles may be electricallydetected with high sensitivity without the use of fluorophores or otherlabels. The examples and embodiments discussed herein may be describedwith reference to the detection of either particles or molecules andsuch terms are used interchangeably herein.

The horizontal nano-sensors may be fabricated using a simplifiedmanufacturing flow having reduced complexity and increased throughput incomparison to known nanoneedle fabrication methodologies. Due to thesimplified flow, a cost effective on-die nano-sensor array may beimplemented with a large number of nano-sensors (on the order ofthousands, tens of thousands, or millions).

In one or more embodiments, the nano-sensor array may implementimpedance detection circuitry on-chip alongside a large number ofnano-sensors, which are distributed along various locations within oradjacent to a fluidic channel. By increasing the number of nano-sensors,a large sample area may be tested simultaneously, resulting in reducedtesting time, increased sensitivity, and ability to perform a largernumber of different tests with reduced reagent usage.

In some embodiments, different tests may be performed by differentnanoneedles, by binding different probes to different nanoneedles.Instead of attaching probes simultaneously to all nanoneedle activeareas, which may include the nanoneedles and/or the dielectric betweenthe nanoneedles, a photoactivatable moiety may be bound to the activeareas of said nanoneedles. Subsequently, in a step which may be repeatedas many times as needed, to bind as many different probes as may bedesired, different photoactivatable moieties may be activated by, forexample, a laser, prior to or concurrently with the introduction of aprobe, which may have a linker moiety which may bind to said activatedmoiety.

FIG. 1 shows a 3-dimensional view of a nano-sensor array having aplurality of horizontal nano-sensors. The nano-sensor array 100 includesbase 102 with a channel 106.

The base may consist solely of a substrate layer or may comprise asubstrate having one or more layers of material formed on the substrate.A plurality of nano-sensors 104 are formed horizontally on the base 102overhanging the channel 106. The nano-sensors 104 are each configured toinclude a probe or probes 112 on a first end of the nano-sensors 104located overhanging the channel 106. The probe or probes is/are designedto bind with a particular type of particle to be detected. Differentnano-sensors may be configured with different probes to allow differenttypes of particles to be detected in the same sample. Different probesmay have different shapes. For ease of illustration and description,probes may be depicted herein as Y forked segments, which are notintended to provide an accurate representation of an actual probe.

During operation, a sample to be tested is passed through the channel.The sample may be presented at an edge of the channel 106 and drawn intothe channel via pressure and/or capillary forces or may be otherwisedelivered into the channel by other means. As particles in the sample108 pass through the channel and interact with the sensor, particlesthat are the target type to be detected bind with the probes 112. Thehorizontal nano-sensors exhibit a change in impedance as a result of atarget particle 108 binding with the probe 112 attached to the end ofthe nano-sensor 104.

The impedance may be measured across two terminals of the nano-sensor104 as depicted by impedance graph 110 using known impedance detectiontechniques. For example, impedance can be determined by measuring thesmall signal AC currents and voltages. For the input, the voltage ismeasured across the input terminals and the current measured byinserting the meter in series with the signal generator. One may use afixed frequency, for example 1 kHz, and set the generator level to, forexample, around 20 mV RMS. For example, in the case where one applies 20mV RMS and measures 10 uA for current, then the impedance is 2 k. Withhigh impedance circuits, the current will become very small anddifficult to measure, so an alternative method is called for, such asusing a fixed resistor and measuring AC voltage at points across theresistor utilizing for example, a trans-impedance amplifier, accordingto known principles of circuit analysis, or using other known methods toamplify the current prior to measurement of said current.

For clarity and ease of illustration, the embodiments described hereinare primarily illustrated and described with reference to an uncoveredchannel formed in a base. It is recognized the embodiments are primarilyenvisioned as having a covered channel but may alternatively have anuncovered channel in some embodiments.. The cover may also include afluidic channel formed on the bottom of the cover and aligned with thefluidic channel of the base. It is recognized that the cover appliedover the channel may be composed of any of a variety of materialsincluding PDMS, Glass, Plastic, Silicon, etc.

FIGS. 2 a-2 d show top and cross sectional views of a nano-sensor arraysimilar to that shown in FIG. 1. FIG. 2 a shows a top view of a secondnano-sensor array having a plurality of horizontal nano-sensors; FIG. 2b shows cross section 1 of the nano-sensor array shown in FIG. 2 a; FIG.2 c shows cross section 2 of the nano-sensor array shown in FIG. 2 a;and FIG. 2 d shows cross section 3 of the nano-sensor array shown inFIG. 2 a. The nano-sensor array 200 includes a base 202 with a channel206 formed in the base. A plurality of nano-sensors 204 are formedhorizontally on the base 202 and overhang a portion of the channel 206.

As described with reference to the nano-sensor shown in FIG. 1, thenano-sensors 204 are each configured to include a probe 212 on a firstend of the nano-sensors 204, which overhangs the channel 206 (probe 212not shown in cross sections). During operation, a sample to be testedpasses through the channel and particles that are the target type (notshown) bind with the probes upon contact.

The horizontal nano-sensors 204 are implemented with a first nanoneedle220, a dielectric 222, and second nanoneedle 224 arranged in a verticalstack with the dielectric 222 separating the first and secondnanoneedles. The lower nanoneedle 220 is formed along a surface of thebase 202. In this illustrated implementation, the dielectric 222 is alsoformed between adjacent ones of the nano-sensors 204, and provides avertical extension to the sidewalls of the channel 206. It is understoodthat additional layer(s) may also be formed over the nano-sensors orbetween the nano-sensors and the base. In this example, probes 212 areaffixed to dielectric 222 at the end of the nano-sensor overhanging thechannel 206. Alternatively, the probe may be affixed to the first and/orsecond nanoneedles 220 and 224, or may be provided on the first and/orsecond nanoneedle and on the dielectric between the first and secondnanoneedle.

In some embodiments, an optional passivation or oxidation layer(s) (notshown) may be formed to cover various portioned of the nanoneedles toreduce the surface area of the nanoneedle which may be available forbinding or interaction with the fluid.

The horizontal nano-sensors 204 each exhibit an impedance betweennanoneedles 220 and 224 that is related to a number of target particlesbound with the probe 212 attached to the end of the nano-sensor 204. Inthis illustrated implementation, a second end of each of the nanoneedles220 and 224 is left exposed and may be used as terminals to measureimpedance between the nanoneedles of the nano-sensor. In someimplementations, contact or bonding pads (not shown) may also be formedon the exposed portion of the second ends.

It is recognized that detectable changes in impedance may includeparasitic impedances from sources other than the desired impedanceassociated with nano-sensors 204. Parasitic impedances reduce the signalto noise ratio of detectable impedance from the nano-sensors 204 andtherefore decrease the sensitivity of detection.

The horizontal implementation of the nanoneedles on the base allowsimpedance detection circuits to be integrated onto the same die in closeproximity to the sensing tip of nano-sensors. In this manner, the lengthof the signal path between the sensing tip of a nano-sensor and theimpedance detection circuit can be reduced to limit the introduction ofexternal noise sources.

The nano-sensor arrays illustrated in FIG. 1 and FIGS. 2 a-2 d may befabricated using the following general method. A base is provided. Thebase may consist solely of a substrate layer or may comprise a substratehaving one or more layers of material formed on a substrate. A pluralityof horizontal nano-sensors is provided along a top surface of the body.Each nano-sensor is fabricated by forming a first nanoneedle along asurface of the base, such that the first nanoneedle has a first endadjacent to or within a region. A dielectric on the first nanoneedle anda second nanoneedle is formed on the dielectric layer. The secondnanoneedle is separated from the first nanoneedle by the dielectric andhas a first end adjacent the first end of the first nanoneedle.

After forming the plurality of nano-sensors, a fluidic channel is etchedin the base within the aforementioned region. In some embodiments, thechannel is etched so that a portion of the fluidic channel will beunderneath of the portion the nano-sensor that will eventually overhangthe channel. After formation of the channel in the base, a cover isprovided and aligned over the channel. In some embodiments, the covermay include an inverted channel formed on the bottom of the cover andaligned with the channel formed in the base. After alignment, the coveris bonded on top of the nanoneedle devices to provide a covered channel.

To assist in the detection of particles of a select target type, aselected probe molecule may be affixed at a sensing tip of thenanoneedles. The selected probe is configured to bind to particles ofthe target type that contact the probe. The structures used to formdifferent probes may have different shapes and may be biological ornon-biological depending on the type of molecule that is desired to bedetected and may be affixed to the sensor using various known bindingmethods. Depending on the application, probe molecules may be affixedduring manufacture of the nano-sensor, in a downstream manufacturingprocess, or by an end-user of a device containing the nano-sensor.

In some embodiments the nanoneedles may have an exposed surface which iscomposed of gold; the probe may have a thiol moiety bound to said probe.The probe may be introduced into the fluidic channel, whereby the probemay thence bind to the exposed gold surface of the nanoneedle, forming apartially covalent gold thiol bond in a thiolate-metal complex, whichmay be preferentially bound to the exposed portions of nanoneedles insaid fluidic channel, and not in other locations in said fluidicchannel. In other embodiments, the dielectric may be fabricated of amaterial which may be silanized. In such an embodiment, the nanoneedleand surrounding areas may be covered with a resist which is also usefulto block an organofunctional alkoxysilale, prior to etching the tip ofthe nanoneedle. Said resist may then be utilized to prevent silanizationof other portions of the nanoneedle and/or fluidic channel. The probemay be bound to a silane coupling agent such as a cyanosilane or athiol-terminal silane. The nanoneedles ends may be etched; said probemay subsequently be introduced to the nanoneedle via the fluidic channelwherein the alkoxysilane may interact and bind to the exposeddielectric.

FIG. 3 shows a cross section of one implementation of a horizontalnanoneedle-sensor that may be used to implement a nano-sensor array. Theillustrated nano-sensor is similar to nano-sensor 204 illustrated inFIG. 2 d. The nano sensor is formed along a top surface of base 302 witha portion of the nano-sensor overhanging a channel 324 formed in aregion of the substrate. The nano sensor includes first 310 and second308 conductive nano-needles separated by a dielectric 306. In thisexample implementation, a second dielectric 304 is formed below thefirst nanoneedle 310 and a passivation layer 312 is formed over thesecond nanoneedle stack. Alternately, the dielectric coating can beformed over the nanoneedle surfaces in the channel and then the sensorlocation can be uncovered by local removal of the dielectric.

FIGS. 4 a-4 d illustrate electrical models of the nano-sensorillustrated in FIG. 3. FIG. 4 a shows a circuit diagram of an electricalmodel of the horizontal nanoneedle-sensor shown in FIG. 3. Theresistance of conductive nanoneedle layers is represented by R_(p1) andR_(p2). C_(ox) and C_(firg) correspond to the capacitance of dielectriclayers and fringing capacitance respectively. The exposed end portion ofthe nanoneedle experiences two double layer capacitances (C_(dl1) andC_(dl2)) between electrode interfaces, formed at endpoints C and D, whenan ionic solution is present. Double layer capacitance depends on theionic concentration and metallic (conductive) surface area [e.g. 0.20uF/mm2 for PBS 50 mM]. Ionic resistance of the solution is shown withRpbs, which varies with the type and concentration of ionic solution,bio-molecule concentration, the geometry of the fluidic channel, and thethickness of middle oxide layer. The sensitivity of the nanoneedle maybe tuned to better detect the double layer capacitance (C_(dl1) andC_(dl2)), or the resistance associated with the sensing region(R_(pbs)), by selecting the frequency of operation, permitting detectionof different moieties with different binding conditions andconductivities.

Operation of the nano-sensor may include two or more phases which couldinclude: a loading phase and a detection phase. In the loading phase,the probe is immobilized by the nano-sensor. The capturing and bindingof target bio-molecule with the probe molecule is called detectionphase. The following examples are describe with reference to these twophases for ease of description. It is understood that operation of thenano-sensor may include additional phases as well, including: wash stepsto remove bound target, incubation time interval to permit time forbinding, etc.

FIG. 4 b shows a circuit diagram of an electrical model of the impedancebetween endpoints A and B of the horizontal nanoneedle-sensor shown inFIG. 3. This model illustrates the modulation of impedance that resultsfrom a target molecule binding with the sensor. Depending on the number,conductivity and size of target molecules bound to the sensor, and thebulk resistivity of the reagent solution, the nano-sensor will exhibit aresistance and capacitance between endpoints A and B related to thenumber of particles bound to the nano-sensor. FIGS. 4 c and 4 dillustrate two binding scenarios that may occur depending on the size ofmolecules and ionic concentration of the sample. FIG. 4 c shows acircuit diagram illustrating the electrical behavior of the horizontalnanoneedle-sensor shown in FIG. 3 resulting from a first particlebinding scenario. In this scenario, a target molecule may bind to thesurface of nanoneedle at the double-layer area 320. As a result of thebinding of the target molecule(s), ions are relocated and shield thesurface of the electrode (C_(dl1), C_(dl2)). In some embodiments, targetmolecules may not bind directly to the surface of the nanoneedle, or toprobes attached thereto, but may instead bind or react within thesensing region of the nanoneedle, for example, on a bead or particle 322which is positioned, held or bound such that the bead or a portionthereof is held within the sensing region of the nanoneedle. FIG. 4 dshows a circuit diagram illustrating the electrical behavior of thehorizontal nanoneedle-sensor shown in FIG. 3 resulting from a secondparticle binding scenario. In this second scenario, which may be morecommon in some biosensing applications, the target molecules may bind tothe sensor outside of a shielded layer from the needle's surface atlocations such as 322. In this scenario, the impedance is changed as aresult of a target molecule being in close proximity to the nano-sensor.

It is recognized that a target molecule may bind to either the surfaceof the nano-sensor (loading phase) or to the probe molecule affixed tothe sensor (detection phase). In both cases, in addition to the changeof double-layer capacitance, ionic conductivity of the solution maychange due to change of local ionic concentration of the solution aroundthe needle's surface. As a result of depletion or accumulation ofcations or anions, the Rpbs is changed. The resistive modulation istypically dominant in comparison to the double layer capacitance. Thisis true if the entire solution (the bulk) has its conductivity changed.If conductivity is locally changed, the conductivity will change onlybriefly.

The above electrical models and description thereof are provided forillustrative purposes and are not intended to represent fully accuratemodels of the impedance modulation or prescribe to any particular theoryof operation.

The parasitic capacitance of the shank of the needle and the connectionto the sense amplifier may limit the sensitivity of the nano-sensor. Itis recognized that decreasing the capacitance C_(ox) in the model shownin FIG. 4 a should cause the poles to separate from each other andcreate a wider frequency range for impedance detection. In one or moreembodiments, the thickness of the dielectric 306 between the nanoneedles308 and 310 is increased while maintaining a thin dielectric layer 306at the sensor tip between endpoints C and D. In one or more embodimentsthe dielectric 306 separating the nanoneedles has a graduated thickness,being thinner at a first end than at a second end.

The detection limit or the minimum detectable concentration ofbio-species in an injected media is a figure of merit in biosensors,which depends in part on the sensitivity (the minimum number of similarbinding events to occur to get a detectable signal), the flow rate,diffusion time of target molecules, sensor geometry, binding kinetics,concentrating field effects, input concentration, inhibitory factors,depletion of input concentration levels, volume of reaction chamber,etc. Reagents may also be added to the solution to increase theconductivity of the sample. The horizontal nano-sensor exhibits a highsensitivity because impedance is modulated at a detectable level inresponse to a small number of binding events in the nanogap sensing areabetween the nanoneedles of the nano-sensor. Because fewer targetmolecules are required for the detection, the presence of targetmolecules may be detected at very low concentrations on the order of 500aM.

The examples herein are primarily illustrated and described withreference to nano-sensors including a nanoneedle/dielectric/nanoneedlearrangement formed in a vertical stack on a substrate base. It isunderstood that the horizontal nanoneedles may also be implemented in anumber of other configurations as well. For example, FIGS. 5 a-5 d showan alternate embodiment of the structure shown in FIG. 1, where bothnanoneedles of each sensor are formed along the surface of the base.FIG. 5 a shows a top view of another nano-sensor array having aplurality of horizontal nano-sensors; FIG. 5 b shows cross section 4 ofthe nano-sensor array shown in FIG. 5 a; FIG. 5 c shows cross section 5of the nano-sensor array shown in FIG. 5 a; and FIG. 5 d shows crosssection 6 of the nano-sensor array shown in FIG. 5 a. Similar to thenano-sensor array 200 shown in FIGS. 2 a-2 d, nano-sensor array 500includes base 502 with a channel 506 formed in the substrate. The basemay consist solely of a substrate layer or may comprise a substratehaving one or more layers of material formed on the substrate.

A plurality of nano-sensors 504 is formed horizontally on the base 502,wherein each nano-sensor may overhang a portion of the channel 506. Eachnano-sensor 504 is also configured to include a probe 512 on a portionof the nano-sensors 504 that overhangs the channel 506 (probe 512 notshown in cross sections). During operation, a sample to be tested passesinto the channel and particles that are the target type (not shown) bindwith the probes upon contact.

The horizontal nano-sensors 504 are implemented with a first nanoneedle520, a dielectric 522, and second nanoneedle 524 arranged adjacent toone another in the same layer above the base 502, with the dielectric522 separating the first 520 and second 524 nanoneedles. In this exampleimplementation, both nanoneedles of each nano-sensor 504 are formedalong a surface of the base 502. In this illustrated implementation, thedielectric 526 is formed in between and over the nano-sensors 504, andprovides a vertical extension to the sidewalls of the channel 506. Likethe nanoneedles sensors 204 illustrated in FIGS. 2 a-2 d, a second endof each of the nanoneedles 520 and 524 is left exposed and may be usedas terminals to measure impedance between the nanoneedles and may alsoform a base for contact pads (not shown). In some embodiments, anoptional passivation or oxidation layer(s) (not shown) may be formed tocover various portions of the nanoneedles to reduce the surface area ofthe nanoneedle which may be available for binding or interaction withthe fluid.

Current biosensors are implemented in a manner that restricts diffusionand/or detection of target molecules to a two-dimensional plane, such ason an optical slide. It is recognized that the channel may beimplemented to any of a variety of different depths. In one or moreembodiments, the channel may be implemented with a depth that allowsparticles to diffuse in a 3-dimensional space. Due to the suspendedgeometry of the needle in the channel, diffusion takes place in threedimensions, which results in a higher rate of binding of targetmolecules to the probe molecule/sensor, and thus a faster detectionplatform.

The examples herein are primarily illustrated and described withreference to nano-sensors overhanging the channel formed in the base. Itis envisioned that the sensors may be implemented overhanging thechannel, flush with the walls of the channel, or any combinationthereof. FIG. 6 illustrates a cross-section view of an alternativeembodiment of the nanoneedle structure shown in FIG. 2 d, in which endsof nano-sensors 204 are flush with the wall of a channel formed in thebase. In yet some other embodiments, the plurality of nano-sensors maybe implemented such that different ones of the nano-sensors overhang thechannel by different lengths, or that some overhang, and others do not.The variation of the overhang length increases the distribution of thesensors in the channel, which is likely to increase target binding rate.Similarly, different sensors may be implemented to be located atdifferent heights.

FIG. 7 illustrates a cross-section view of an alternative embodiment ofthe nanoneedle structure shown in FIG. 2 d, wherein the channel isimplemented by sidewalls formed above the substrate. In thisillustration, the channel 206 is not formed in the base 202. The channel206 is formed by depositing material 702 on the substrate and over thenano-sensors 204 to form sidewalls of the channel 206.

FIG. 8 illustrates a top view of double and quad nanoneedles. On theleft, two of the nano-sensors 802 and 810 are formed on a body 806 asdiscussed above. On the right, four nano-sensors 812, 816, 826, and 828are formed on a substrate.

In order to minimize parasitic capacitance, nanoneedles may befabricated such that they have thin regions of dielectric close to thesensing regions, and thicker regions farther from the sensing region.These differences in the thickness of different portions of thenanoneedles may be formed by applying a first nanoneedle, followed by auniform layer of dielectric applied over said first nanoneedle. An etchresist may then be applied over portions of the nanoneedle, and may alsobe applied over portions of the adjacent silicon. Thinner portions ofnanoneedles 802 and 810 are thinner portions of the dual nanoneedles,while thinner portions of nanoneedles 812, 816, 826 and 828 are thinnerportions of quad nanoneedles, formed by etching, including etchingregions of silicon adjacent to dual nanoneedle 806, and regions adjacentto quad nanoneedle 824; Those portions of the nanoneedles which are notetched and which are illustrated include thicker portion of dualnanoneedle 818 and thicker portions of nanoneedles 818, 820.

The nanoneedles of the nano-sensors may be formed using any one of anumber of lithography techniques. One method suitable for formation ofthe horizontal nanoneedles is known as electron-beam lithography (EBL).In EBL, a resist layer is formed on a silicon substrate. The resistlayer is exposed with a beam of electrons in a patterned fashion acrossthe surface of the resist layer. Following patterning, the resist layeris developed by selectively removing either exposed or non-exposedregions of the resist layer to form patterned channels in the resistlayer, which may form a mask for the nanoneedles. Depending on thematerials used, the nanoneedles may be formed using a number of methodsrecognized by those skilled in the art e.g. electroplating, chemicalvapor deposition (CVD), etc. In another example implementation, an etchstep may be performed to etch a conductive layer below the patternedresist layer to form the nanoneedles. Other possible patterning andetching methods are envisioned as well. EBL is not limited to thediffraction limit of light and allows photo-patterning techniques tocreate features in the nanometer range. It is understood that the EBLprocess may be implemented using a number of different configurationsutilizing different electron beam energies, various resist materials,and various chemical etch techniques.

In one example EBL implementation, a silicon substrate is cleaned,coated with 100 nm thick 2% 950K Poly Methyl Methacrylate positive toneelectron beam resist, and soft baked for 120 s at 200 ° C. An electronbeam having at an acceleration voltage of 10 kV and an electron dose of100 μC/cm2 may be used to pattern the resist layer. After EBL exposure,the resist is developed by soaking in a methyl isobutyl ketone/isopropylalcohol (MIBK/IPA) 1:3 by volume solution for 30 s at 18 ° C., and thenrinsed with pure 2-propanol for 30 seconds. It is recognized that thewidth of the patterned channels, and the resulting nanoneedles, varieswith different dose factors (e.g. Single Pixel Lines (SPL) measuring 20nm at SPL dose =345 pC/cm). The minimum width of continuous etched SPLwas measured as 8.93 nm (FIG. 7, d) but it results in high LER (lineedge roughness).

The nanoneedles may be formed using a number of different conductivematerials including, e.g., p+-silicon, Al, Au, etc. For ease ofdescription, the examples herein are primarily described with referenceto nanoneedles formed using p+ silicon as the conductive material.

The following example describes one possible implementation formanufacturing nanoneedles with p+ silicon. 250 nm of silicon oxide isthermally grown on the silicon substrate, followed by the deposition of100 nm of poly-silicon, then doped with phosphorus to achieve a sheetresistance of 210 ohms per square. (It was reduced to 30 ohms per squareusing alternative doping approaches.) This p+-silicon layer is coatedwith 30 nm of SiO2 followed by another p+-silicon layer deposition.Finally, a 20 nm SiO2 layer covers the top p+-silicon layer. A threestep lithography process is performed, followed by etching thatincludes: a 1.6 um SHIPLEY 3612 resist spin-coated (SVG coat), and afterlithography (using a Karl Suss mask aligner) and baking for 120 sec, theresist is baked and developed (SGV Photoresist Developer). Then thecombination of the 20 nm top-SiO2 layer, the 100 nm top p+-siliconlayer, and the 30 nm middle oxide layer down to the bottom p+-siliconlayer, are dry-etched (Applied Materials Precision 5000 Etch). In thesecond lithography step, the remaining 100 nm bottom p+-silicon layerand 200 nm bottom SiO2 layer as well as 1 um deep in the substratesilicon is dry-etched followed by a XeF2 injection to etch and open thechannel underneath of the nanoneedle devices. Finally, the 20 nm topsilicon layer is removed from the pad areas to enable contact access formeasurement and testing.

The manufacturing implementation using p+ silicon to form nano-wiresdescribed above is provided for illustrative purposes only. Thethicknesses, selection of materials, and other process specifics areintended only for illustration and not limitation.

The embodiments are thought to be applicable to a variety ofapplications, which utilize particle detection. For example, on-chipamplification of electrical signals and impedance detection circuits maybe integrated alongside the nano-sensor arrays to provide lab-on-a-chipdiagnostics for portable devices and are thought to have useful clinicalapplications for detection of several biomarkers in early stagediagnoses, and in the field diagnostics. Other aspects and embodimentswill be apparent to those skilled in the art from consideration of thespecification. It is intended that the specification and illustratedembodiments be considered as examples only, with a true scope and spiritof the invention being indicated by the following claims.

What is claimed is:
 1. A method for the detection of target chemical andbiological particles comprising: providing a sensing device comprising abase and a plurality of nano-sensors in a fluidic channel, each of saidplurality of nano-sensors comprising a first nanoneedle and a secondnanoneedle, wherein the first and second nanoneedles are separated by asolid state dielectric and have ends that are adjacent to one another;passing target particles through said fluidic channel with the aid offluidic pressure; and sensing the presence of said target particlesproximate to the plurality of nano-sensors by detecting a change inelectric signals through the nanoneedles.
 2. The method of claim 1,wherein the plurality of nano-sensors further comprise a probe proximateto an exposed portion of the solid state dielectric between the firstand second nanoneedles at a first end of the second nanoneedle, theprobe configured and arranged to bind to said target particles.
 3. Themethod of claim 2, wherein the probe is provided on one of the first andsecond nanoneedles.
 4. The method of claim 2, wherein the sensing devicefurther comprises a cover over said channel, said cover comprising aninverted fluidic channel.
 5. The method of claim 2, wherein the solidstate dielectric is thicker at one portion of the first and secondnanoneedles than at a first end of said nanoneedles.
 6. The method ofclaim 2, wherein the second nanoneedle is formed adjacent to a surfaceof the base.
 7. The method of claim 2, wherein the second nanoneedle isdisposed vertically in relation to the first nanoneedle.
 8. The methodof claim 2, wherein the first and second nanoneedles are disposedvertically in relation to the base.
 9. The method of claim 2, whereinthe target particles are polynucleotides.
 10. The method of claim 3,wherein the change in said electric signals is a change in impedancebetween the first and second nanoneedles in response to target particlesbeing bound to the probe.
 11. The method of claim 3, wherein the sensingdevice further comprises a passivation layer over at least the first andsecond nanoneedles, the passivation layer not covering at least aportion of a first end of the first nanoneedle or a first end of thesecond nanoneedle.
 12. A nano-sensor array, comprising: a sensing devicecomprising a base; a channel proximate to the base wherein particles arepassed through said channel via fluidic pressure; plurality ofnano-sensors wherein each sensor comprises: nanoneedle and a secondnanoneedle, wherein the first and second nanoneedles are separated by asolid state dielectric and have ends that are adjacent to one another;and a detection circuit coupled to each of said nanoneedles, wherein thedetection circuit detects a change in impedance between the first andsecond nanoneedles of each nano-sensor that results from the binding ofa target particle to a respective probe.
 13. The nano-sensor array ofclaim 12, wherein said probe is provided on one of the first and secondnanoneedles.
 14. The nano-sensor array of claim 12, further comprising asecond particle bound to at least one said particle of a target type.15. The nano-sensor array of claim 13, wherein the detection circuitdetermines the number of target particles bound to the respective probe.16. The nano-sensor array of claim 13, wherein each probe in a firstsubset of the plurality of nano-sensors binds to a first type of targetparticle and each probe in a second subset of the plurality ofnano-sensors binds to a second type of target particle.
 17. Thenano-sensor array of claim 13, wherein the second nanoneedle of eachnano-sensor is disposed along the surface of the base.
 18. Thenano-sensor array of claim 13, wherein the second nanoneedle of eachnano-sensor is disposed vertically in relation to the first nanoneedle.19. The nano-sensor array of claim 13, wherein the first and secondnanoneedles are disposed vertically in relation to the base.
 20. Thenano-sensor array of claim 13, wherein the solid state dielectric has athickness of 500 nm or less.
 21. The nano-sensor array of claim 14,wherein the second particle is a bead.
 22. A device detecting ormeasuring a biological reaction, the device comprising a base with aplurality of nano-sensors, each nano-sensor comprising two nanoneedlesand a dielectric layer between the two nanoneedles, the device beingadapted to measure an electric signal resulting from the biologicalreaction, the nanoneedles further comprising a section that is designedto bind to a biological particle.